Multiple detection based on lateral flow immunoassay (LFA) holds significant research importance across various fields. Recently, the use of multi-color surface-enhanced Raman scattering (SERS) nanotags has enabled simultaneous detection of multiple targets within a single test line (T-line) in LFA, greatly enhancing detection efficiency, reliability, and application flexibility. However, challenges regarding signal overlap in the Raman-fingerprint region (400–1800 cm) and the difficulty in determining specific targets based on typical color analysis have persisted. In this study, we developed two types of nanoprobes that encode both color and Raman information to address these challenges in simultaneous multiple detection on a single T-line in SERS-LFA. The nanoprobes were encoded with distinguishable blue and red colors, respectively, enabling colorimetric study in multiplex LFA for qualitative analysis. Additionally, they were encoded with distinctive and non-overlapping Raman signals in the Raman-silent region (1800–2800 cm), ensuring accurate and sensitive detection by effectively avoiding interference from each other as well as background signals in the sample matrix. Using carbendazim (CBZ) and imidacloprid (IMI) as model targets, the color- and Raman-encoded SERS-LFA demonstrated promising potential for the simultaneous detection of CBZ and IMI in both standard and real samples, whether in a colorimetric or a SERS mode.

中文翻译：

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无颜色和背景的拉曼编码侧流免疫分析，用于在单条测试线中同时检测多菌灵和吡虫啉

基于侧流免疫分析 (LFA) 的多重检测在各个领域具有重要的研究重要性。最近，使用多色表面增强拉曼散射（SERS）纳米标签实现了LFA中单条测试线（T线）内的多个目标的同时检测，大大提高了检测效率、可靠性和应用灵活性。然而，拉曼指纹区域（400-1800 cm）信号重叠以及基于典型颜色分析确定特定目标的困难仍然存在。在这项研究中，我们开发了两种类型的纳米探针，可编码颜色和拉曼信息，以解决 SERS-LFA 中单条 T 线同时多重检测的挑战。纳米探针分别用可区分的蓝色和红色进行编码，从而能够在多重 LFA 中进行比色研究以进行定性分析。此外，它们在拉曼沉默区域（1800-2800 cm）内用独特且不重叠的拉曼信号进行编码，通过有效避免相互干扰以及样品基质中的背景信号来确保准确和灵敏的检测。使用多菌灵 (CBZ) 和吡虫啉 (IMI) 作为模型目标，颜色和拉曼编码的 SERS-LFA 显示出在标准样品和实际样品中同时检测 CBZ 和 IMI 的巨大潜力，无论是在比色还是 SERS 模式下。